Isolation, characterization, and identification of proteins interfering with enzyme-linked immunosorbent assay of antibiotics in fish matrix

Abstract

The interaction between some proteins and immune globulin has been confirmed as an important source of matrix interference with the immunoassay of fishery products, but detailed biochemical properties of these proteins have not been indicated. Two interference-inducing proteins (42 and 36 kD) in flounder were isolated, characterized, and identified. Their influences on the immunoassay of norfloxacin were confirmed by western blotting and enzyme-linked immunosorbent assay. The pI value and pH stability of the two proteins were also investigated. Using LC-MS/MS, the two proteins were identified as fructose-bisphosphate aldolase, and such results were partly verified by the aldolase activity of the 42 and 36 kD isolates. Considering the prevalence of these proteins (as multi-functional aldolase of muscles) in foods, these results would help to further understand the matrix effects in various immunoassays as well as the development of effective techniques to improve the efficiency of immunoassays.

The effect of chlorophyll on the enzyme‐linked immunosorbent assay (ELISA) of procymidone in vegetables and the way to overcome the matrix interference

“…Chlorophyll is unstable and easy to decompose under acidic conditions, 31 therefore a new idea based on the pretreatment with acids was suggested here. Among various acids, 5‐sulfosalicylic acid has demonstrated excellent ability to eliminate matrix proteins with little effect on the ELISA and other immunoassays 24,32,33 . Considering the excellent stability of procymidone under acidic conditions, in principle 5‐sulfosalicylic acid would not react with procymidone or lead to its breakdown 34 .…”

“…Among various acids, 5-sulfosalicylic acid has demonstrated excellent ability to eliminate matrix proteins with little effect on the ELISA and other immunoassays. 24,32,33 Considering the excellent stability of procymidone under acidic conditions, in principle 5-sulfosalicylic acid would not react with procymidone or lead to its breakdown. 34 These proposals were confirmed well by the following experiments.…”

Heterologous Expression, Characterization, and Fish Matrix Performance of a Shark-Derived Single-Domain Antibody Fusion Protein against Enrofloxacin

“…Previous researches have demonstrated that the matrix components, notably proteins in sh and other animal-derived foods, might create signi cant matrix effects in immunoassays since the accuracy of immunoassays can be impacted by sample matrix (Wang et al 2016;Wang et al 2013;Xu et al 2012). As can be seen from Fig.…”